Difference between revisions of "Part:BBa K3895004"
 
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KerBteQ7 is a serine keratinase originally found in ''Bacillus tequilensis Q7''.  
 
KerBteQ7 is a serine keratinase originally found in ''Bacillus tequilensis Q7''.  
 
6x His-tags were added to both sides of the sequence for purification, and constructed into PET-28a(+) (<partinfo>K3895008</partinfo>). The length of KerBteQ7  is 1214 bp.
 
6x His-tags were added to both sides of the sequence for purification, and constructed into PET-28a(+) (<partinfo>K3895008</partinfo>). The length of KerBteQ7  is 1214 bp.
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[[File:T--SZ SHD--Q7O.png|900px|center]]
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|[[File:T--SZ SHD--Q7.jpg|200px|thumb|center|'''Figure 1.''' Plasmid construction of KerBteQ7.]]
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|[[File:T--SZ SHD--Q7.jpg|300px|thumb|center|'''Figure 1.''' Plasmid construction of KerBteQ7.]]
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|[[File:T--SZ SHD--PCR2.png|200px|thumb|center|''' Figure 2.''' PCR and gel electrophoresis of KerBteQ7.]]
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|[[File:T--SZ SHD--PCR2.png|300px|thumb|center|''' Figure 2.''' PCR and gel electrophoresis of KerBteQ7.]]
 
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<partinfo>BBa_K3895004 parameters</partinfo>
 
<partinfo>BBa_K3895004 parameters</partinfo>
 
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===References===
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Zaraî Jaouadi, N., Rekik, H., Ben Elhoul, M., Zohra Rahem, F., Gorgi Hila, C., Slimene Ben Aicha, H., Badis, A., Toumi, A., Bejar, S., & Jaouadi, B. (2015). A novel keratinase from Bacillus tequilensis strain Q7 with promising potential for the leather bating process. International Journal of Biological Macromolecules, 79, 952–964. https://doi.org/10.1016/j.ijbiomac.2015.05.038
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Vidmar, Beti, and Maša Vodovnik. “Microbial Keratinases: Enzymes with Promising Biotechnological Applications.” Food technology and biotechnology vol. 56,3 (2018): 312-328. doi:10.17113/ftb.56.03.18.5658

Latest revision as of 08:08, 11 October 2021


Keratinase kerBteQ7

KerBteQ7 is a serine keratinase originally found in Bacillus tequilensis Q7. 6x His-tags were added to both sides of the sequence for purification, and constructed into PET-28a(+) (BBa_K3895008). The length of KerBteQ7 is 1214 bp.

T--SZ SHD--Q7O.png
Figure 1. Plasmid construction of KerBteQ7.
Figure 2. PCR and gel electrophoresis of KerBteQ7.
Name of keratinase IPTG concentration/mM Temperature/°C Time/h
KerBteQ7 5 16 12

Modeling

The biological activity of a protein is not only determined by the primary structure of the protein molecule but also closely related to its specific spatial structure. Elucidating the process of protein folding in functional and structural details will be of great significance to evaluate their keratinotic functions. According to the amino acid sequences in the enzyme active sites and associated catalytic mechanisms, proteases can be classified into seven broad groups: serine, cysteine, threonine, aspartic and glutamic proteases, metalloproteases and asparagine peptide lyases. According to the nature of their active site, keratinases belong to serine- and metalloproteases or serine metalloproteases. Moreover, the enzymatic degradation of keratin is a multistage process that requires the following steps: adsorption of the keratinases to the surface of macromolecule by electrostatic and hydrophobic interactions, followed by catalytic action [2]. Hence we used Swiss Model to predict their hydrophobic region and serine groups.


Figure 1. Swiss Model showing serine residues of KerBteQ7.
Figure 2. Swiss Model showing hydrophobisity of KerBteQ7 .
Figure 3. Ramachandran Plot of KerBteQ7.Q7 Ramachandran Favoured is 91.25%

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 457
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1156


References

Zaraî Jaouadi, N., Rekik, H., Ben Elhoul, M., Zohra Rahem, F., Gorgi Hila, C., Slimene Ben Aicha, H., Badis, A., Toumi, A., Bejar, S., & Jaouadi, B. (2015). A novel keratinase from Bacillus tequilensis strain Q7 with promising potential for the leather bating process. International Journal of Biological Macromolecules, 79, 952–964. https://doi.org/10.1016/j.ijbiomac.2015.05.038

Vidmar, Beti, and Maša Vodovnik. “Microbial Keratinases: Enzymes with Promising Biotechnological Applications.” Food technology and biotechnology vol. 56,3 (2018): 312-328. doi:10.17113/ftb.56.03.18.5658